ABSTRACT The significant challenge underlying metabolomics studies is that lack of annotation for the vast chemical space despite efforts to catalogue the human metabolome and to develop more comprehensive mass spectrometry- based libraries of endogenous and xenobiotic compounds. Consequently, application of high resolution and high mass accuracy mass spectrometry is critical to the interpretation of unknowns, and the unique features (high resolution, MSn) of the Thermo Scientific Orbitrap Fusion Lumos Tribrid mass spectrometer will be integral toward structural elucidation of these unknowns. Acquisition of the new Thermo Scientific Orbitrap Fusion Lumos Tribrid will provide both additional capacity and capabilities for The Pennsylvania State University Metabolomics Facility. The projects described herein will receive priority access to the new instrument and occupy the majority of the AUT. While off-loading these project samples to the new instruments will free up some service hours from existing instruments, some of these project refinements will generate additional sample runs to fully exploit the enhanced potential for characterizing the metabolome and biochemical pathways. However, the Metabolomics Facility also has a backlog of sample run requests from new users and for new projects, and these will rapidly fill in any runtime on other instruments. We describe in detail nine projects that require the unique capabilities of the Thermo Scientific Orbitrap Fusion Lumos Tribrid. The new capabilities of this instrument are essential for the success of NIH-funded investigators as the currently available instrumentation both on campus and in neighboring institutions is not sufficient in terms of sensitivity and resolution to complete the proposed studies. The instrument will be housed in the state-of-the-art metabolomics facility at The Pennsylvania State University and will be invaluable not only as a research tool but also a means for educating undergraduates, graduate students, postdoctoral scholars, and investigators throughout the university. Overall The Pennsylvania State University has made a strong commitment to support this new platform by supporting its operation for seven years, fully funding the salary of a research associate for two years, and supporting the salary of the facility director.